Method And Compositions For Improving Selective Catabolysis And Viability In Cells Of Keratin Surfaces

ABSTRACT

A composition for treating keratin surfaces to stimulate selective catabolysis and improve cellular viability comprising at least one autophagy activator and at least one DNA repair enzyme, and a method for improving selective catabolysis and cellular viability by treating with the composition.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority from U.S. ProvisionalApplication No. 61/749,633, filed Jan. 7, 2013.

TECHNICAL FIELD

The invention is in the field of compositions for application to skinwhich enhance selective catabolysis in skin cells.

BACKGROUND OF THE INVENTION

It is well known that stress and environmental aggressors such as UVlight, pollution, and cigarette smoke can be very detrimental to skinand accelerate the appearance of aging. Exposure to stress andenvironmental aggressors often causes damage to cellular DNA,mitochondria, and cellular proteins, lipids, and tissue. Damagedcellular material found within the cell, for example, cytoplasmic ororganelle debris can exert a toxic effect on cells by impeding theirnormal metabolic processes.

Healthy cells have a normal cleansing process that eliminates damagedcellular material and debris. Such detoxification often occurs through aphagocytic process referred to as autophagy, where the cellular debrisis engulfed within a vacuole and degraded with cellular enzymes such aslysozymes. Autophagy and the mechanism of autophagy activation in skincells is described in U.S. Patent Publication No. 2011/0243983 A1,hereby incorporated by reference in its entirety. There is much interestin formulating skin treatment compositions to contain ingredients thatstimulate cellular autophagy because the ability of cells to cleanse anddetoxify themselves of debris that otherwise impedes healthy metabolicfunction is improved. The cleansing through autophagy creates newsources of energy for cellular functions because the degradationproducts release building blocks such as amino acids that can berecycled by the cell. Improved cellular metabolic function in turn meansimproved cellular health and longevity and greater resistance toundesirable side effects of aging such as lines, wrinkles, mottled skin,hyperpigmentation, laxity and so on.

DNA repair enzymes are known to repair DNA in various ways. Such enzymesare generally categorized according to the type of DNA base mutagenicdamage that is repaired.

It has been discovered that the combination of DNA repair enzymes andautophagy activators most unexpectedly will beneficially affect thenormal cellular cleansing processes, promote cellular health andlongevity, and minimize the deleterious effects of environment, UVlight, pollution, and other environmental insults on skin.

Accordingly it is of interest to maximize the cellular health andlongevity of cells, particularly skin cells such as keratinocytes orfibroblasts, by treating them with ingredients that stimulate naturalcellular repair processes by, for example, eliminating cellular debrisand toxins, and in addition, to maximize the effectiveness of suchcellular detoxification by ensuring that the maximum number ofdetoxification mechanisms are operable. It is also of interest tostimulate detoxification processes where the breakdown of the cellulardebris and toxins results in amino acids or other biological moleculesthat can be recycled by the cell.

Thus, there is a need to maximize cellular health and longevity bystimulating selective catabolysis in cells so that cellular debris andtoxins are removed and by products of such degradation may be recycled.More preferably, the selective catabolysis is due to enhancing autophagyactivity and promoting DNA repair using DNA repair enzymes.

It has been discovered that compositions containing at least oneautophagy activator and at least one DNA repair enzyme in selectivecellular catabolysis, that is, cleansing cells of toxic waste productsby breaking down the waste products into molecules that may be recycledand used by the cell in normal metabolic functions.

SUMMARY OF THE INVENTION

The invention is directed to a composition for treatment of keratinsurfaces to stimulate selective catabolysis comprising at least oneautophagy activator and at least one DNA repair enzyme.

The invention is further directed to a method for stimulating selectivecatabolysis in cells of keratin surfaces by applying to such surfaces acomposition comprising at least one autophagy activator and at least oneDNA repair enzyme.

DETAILED DESCRIPTION I. Definitions

All percentages mentioned herein are percentages by weight unlessotherwise indicated.

All documents mentioned herein are incorporated by reference in theirentirety.

“Autophagy” means the process by which cells cleanse themselves oftoxins and debris by forming a membrane around the debris, segregatingit from the rest of the cell, and adjoining the formed vacuole withcellular lysosomes, which are cellular organelles that contain acidhydrolase enzymes that break down the cellular debris and waste found inthe vacuole.

“Autophagy Activator” means an ingredient that stimulates the normalcellular autophagy processes.

“CLOCK gene activator” means an ingredient that activates one or moreCLOCK genes present in keratinocytes.

The term “DNA repair enzyme” means an enzyme that is operable to repairDNA base mutagenic damage. Such enzymes are often categorized by thetype of DNA damage they repair, for example BER (base excision repair)enzymes, nucleotide excision repair (NER) enzymes; mismatch repair (MMR)enzymes; DNA helicases; DNA polymerases, and so on. For example,mutations such as 8-oxo-7,8-dihydro-2′-deoxyguanosine may be repaired byOGG1 (8-oxoGuanine glycosylase); T-T dimers which may be repaired by(Nucleotide excision repair (NER) Photolyase); 6-4 photoproducts (whichmay be repaired by NER); and 06-methyl guanine (which may be repaired by06-alkyl guanine transferase (AGT)).

The term “keratin surfaces” means skin, hair, and nails.

“PER1 gene activator” means an ingredient that activates one or morePER1 genes found in keratinocytes.

“Proteasome” means a protein complex typically located in the nucleus orcytoplasm of cells that is operable to degrade damaged cellular proteinsby proteolysis into smaller subunits which may then be further digestedinto single amino acids. These recycled amino acids may be used by thecell in the synthesis of new proteins.

“Proteasome activator” means an active ingredient that stimulates theactivity of proteasomes in cells of keratin surfaces such askeratinocytes, fibroblasts, etc.

“Recycle” means, with respect to the degradation of cellular debris andtoxins, that the debris and toxins may be broken down into moleculessuch as proteins, lipids, amino acids, or other biological materialsthat are usable by the cell in its normal healthy metabolic processes.

“Repair” means, with respect to skin cells, that the damaged portions ofcells, such as DNA, mitochondria, proteins, lipids, or other cellularmaterials are reduced or eliminated.

“Selective catabolysis” means, with respect to the cells of keratinsurfaces, that the cells are able to cleanse themselves of debris,waste, and toxins selectively without compromising healthy cellularconstituents, and preferably by one or more of mechanisms such asactivating cellular autophagy or activating cellular proteasomeprocesses.

II. Autophagy Activator

The composition of the invention contains at least one ingredient thatis operable to activate normal cellular autophagic processes. Theautophagy activator is present in amounts ranging from about 0.00001 to20%, preferably 0.0001-15%, more preferably from about 0.001 to 10%. Ingeneral, the cellular autophagy process comprises four general steps.Step 1 is the initiation of vacuole formation; Step 2 the formation ofthe initial vacuole or autophagosome which sequesters the cytoplasmicmaterial to be degraded. Step 3 is the maturation of the autophagosomeinto a degradative vacuole. Step 4 is the actual degradation of thesequestered material.

Ingredients with autophagy activation activity can be identified bytheir ability to either stimulate or inhibit various cellular metabolicpathways. For example, ingredients that stimulate the expression ofMAP-LC3, ATG5-12, protein p53, AMPK, or DRAM are suitable autophagyactivators. Ingredients that inhibit the expression of mTOR are alsosuitable autophagy activators.

The gene MAP-LC3 codes for microtubule-associated protein 1 light chain3, a protein that initiates formation of autophagosomes. ATG5-12 alsostimulates formation of autophagosomes. mTOR, also known as mammaliantarget of rapamycin, is also known as the mechanistic target ofrapamycin or FK506 binding protein 12-rapamycin associated protein 1(FRAP1). FRAP1 is encoded by the FRAP gene. Any ingredient that inhibitsthe expression of mTOR, involved in autophagosome creation, will haveautophagy activating properties. Also suitable as autophagy activatorsare ingredients that stimulate expression of protein p53, AMPK, and/orDRAM (damage remedy autophagy modulator protein) in keratinocytes.Protein p53, also known as a tumor suppressor protein, is encoded by thep53 gene. AMPK means AMP activated protein kinase and DRAM, damagerelated autophagy modulator. Both are known to stimulate autophagyactivation in keratinocytes.

Thus any ingredient that has the above mentioned effects on the genesmay be suitable autophagy activators. During the autophagocytic processcellular debris such as oxidized proteins and peroxidized lipids aredegraded. Such cellular debris often affects normal metabolic function.Screening of ingredients to determine efficacy by ability to stimulateor inhibit cellular, preferably keratinocyte, genes and/or proteinsmentioned above may be done according to methods as set forth in USPatent Publication No. 2011/0243983 or other methods known in the art.

For example, one general process for identifying ingredients that may beautophagy activators is by first inducing nutritive stress in culturedcells such as keratinocytes. For example, the cells are first culturedin complete culture medium with growth factors, for about 24 hours. Theculture medium is then removed and replaced with a non-nutritive culturemedium, for example one that does not contain growth factors. The cellsare cultured for about 30 minutes to about 25 hours in a state ofnutritive stress. Then, the non-nutritive culture medium is removed andreplaced with complete culture medium to promote cellular recovery.Thereafter, the cells are evaluated for autophagocytic activity bymeasuring the expression of one or more of MAP-LC3; ATGS-12;phosphorylated mTOR; phosphorylated p53; DRAM; or phosphorylated AMPK inthose cells. Measurement of such expression can take place byimmunofluorescence measurements. In addition, the expression can beascertained by Western Blot analysis of phosphorylated proteinsassociated with the expressed genes.

Examples of ingredients that are known to exert either the stimulatoryor inhibitory effects on the above mentioned genes which, in turn,stimulate autophagy, are yeast extracts including but not limited tothose from the genuses such as Lithothamnium, Melilot, Citrus, Candida,Lens, Urtica, Carambola, Momordica, Yarrowia, Plumbago, etc. Furtherspecific examples include Lithothamniumn calcaneum, Melilotusofficinalis, Citrus limonum, Candida saitoana, Lens culinaria, Urticadioica, Averrhoa carambola, Momordica charantia, Yarrowia lipolytica,Plumbago zeylanica and so on.

Also suitable are ingredients such as amiodarone hydrochloride, GF109203X which is also referred to as(3-(N-[Dimethylamino]propyl-3-indolyl)-4-(3-indolyl)maleimide3-[1-[3-(Dimethylamino)propyl]1H-indol-3-yl]-4-(1Hindol-3-yl)1H-pyrrole-2,5dione Bisindolylmaleimide I;N-Hexanoyl-D-sphingosine; Niclosamide; Rapamycin from Streptomyceshygroscopicus; Rottlerin which is also referred to as(1-[6-[(3-Acetyl-2,4,6-trihydroxy-5-methylphenyl)methyl]-5,7-dihydroxy-2,2-dimethyl-2H-1-benzopyran-8-yl]-3-phenyl-2-propen-1-one,Mallotoxin); STF-62247, also known as5-Pyridin-4-yl-thiazol-2-yl-m-tolyl-amine; Tamoxifen; Temsirolimus whichis also known as 42-[3-Hydroxy-2-methylpropanoate, CCI-779, Rapamycin;ATG1 autophagy related 1 homolog; ATG1, Serine/threonine-protein kinaseULK1, UNC-51-like kinase; or Z36 which is also referred to as((Z)-5-Fluoro-1-(3′-dimethylamino)propyl-3-[(5′-methoxyindol-3-ylidene)methyl]-indolin-2-one;or1-[3-(dimethylamino)propyl]-5-fluoro-1,3-dihydro-3-[(5-methoxy-1H-indol-3-yl)methylene]-2H-Indol-2-one);Bufalin, also referred to as 3β,14-Dihydroxy-5β,20(22)-bufadienolide,5β,20(22)-Bufadienolide-3β,14-diol. Such ingredients may be purchasedfrom Sigma-Aldrich Chemical Company.

III. DNA Repair Enzyme

The composition contains one or more DNA repair enzymes. Suggestedranges are from about 0.00001 to about 35%, preferably from about0.00005 to about 30%, more preferably from about 0.0001 to about 25% ofone or more DNA repair enzymes.

DNA repair enzymes as disclosed in U.S. Pat. Nos. 5,077,211; 5,190,762;5,272,079; and 5,296,231, all of which are hereby incorporated byreference in their entirety, are suitable for use in the compositionsand method of the invention. One example of such a DNA repair enzyme maybe purchased from AGI/Dermatics under the trade name Roxisomes®, and hasthe INCI name Arabidopsis Thaliana extract. It may be present alone orin admixture with lecithin and water. This DNA repair enzyme is known tobe effective in repairing 8-oxo-Guanine base damage.

Another type of DNA repair enzyme that may be used is one that is knownto be effective in repairing 06-methyl guanine base damage. It is soldby AGI/Dermatics under the tradename Adasomes®, and has the INCI nameLactobacillus ferment, which may be added to the composition of theinvention by itself or in admixture with lecithin and water.

Another type of DNA repair enzyme that may be used is one that is knownto be effective in repairing T-T dimers. The enzymes are present inmixtures of biological or botanical materials. Examples of suchingredients are sold by AGI/Dermatics under the tradenames Ultrasomes®or Photosomes®. Ultrasomes® comprises a mixture of Micrococcus lysate(an end product of the controlled lysis of various species ofmicrococcus), lecithin, and water. Photosomes® comprise a mixture ofplankton extract (which is the extract of marine biomass which includesone or more of the following organisms: thalassoplankton, greenmicro-algae, diatoms, greenish-blue and nitrogen-fixing seaweed), water,and lecithin.

Another type of DNA repair enzyme may be a component of variousinactivated bacterial lysates such as Bifida lysate or Bifida fermentlysate, the latter a lysate from Bifido bacteria which contains themetabolic products and cytoplasmic fractions when Bifido bacteria arecultured, inactivated and then disintegrated. This material has the INCIname Bifida Ferment Lysate.

Other suitable DNA repair enzymes include Endonuclease V, which may beproduced by the denV gene of the bacteriophage T4. Also suitable are T4endonuclease; O⁶-methylguanine-DNA methyltransferases; photolyases suchas uracil- and hypoxanthine-DNA glycosylases; apyrimidinic/apurinicendonucleases; DNA exonucleases, damaged-bases glycosylases (e.g.,3-methyladenine-DNA glycosylase); correndonucleases either alone or incomplexes (e.g., E. coli uvrA/uvrB/uvrC endonuclease complex); APEXnuclease, which is a multi-functional DNA repair enzyme often referredto as “APE”; dihydrofolate reductase; terminal transferase;topoisomerase; O⁶ benzyl guanine; DNA glycosylases.

Other types of suitable DNA repair enzymes may be categorized by thetype of repair facilitated and include BER (base excision repair) or BERfactor enzymes such as uracil-DNA glycosylase (UNG); single strandselective monofunctional uracil DNA glycosylase (SMUG1);3,N(4)-ethenocytosine glycosylase (MBD4); thymine DNA-glycosylase (TDG);A/G-specific adenine DNA glycosylase (MUTYH); 8-oxoguanine DNAglycosylase (OGG1); endonuclease III-like (NTHL1); 3-methyladenine DNAglycosidase (MPG); DNA glycosylase/AP lyase (NEIL1 or 2); APendonuclease (APEX 1 and 2), DNA ligase (LIG3), ligase accessory factor(XRCC1); DNA 5′-kinase/3′-phosphatase (PNKP); ADP-ribosyltransferase(PARP1 or 2).

Another category of DNA repair enzymes includes those that are believedto directly reverse damage such as O⁶—MeG alkyl transferase (MGMT);1-meA dioxygenase (ALKBH2 or ALKBH3).

Yet another category of enzymes operable to repair DNA/proteincrosslinks includes Tyr-DNA phosphodiesterase (TDP1).

Also suitable are MMR (mismatch exision repair) DNA repair enzymes suchas MutS protein homolog (MSH2); mismatch repair protein (MSH3); mutShomolog 4 (MSH4); MutS homolog 5 (MSH5); or G/T mismatch-binding protein(MSH6); DNA mismatch repair protein (PMS1, PMS2, MLH1, MLH3);Postmeiotic segregation increased 2-like protein (PMS2L3); orpostmeiotic segregation increased 2-like 4 pseudogene (PMS2L4).

Also suitable are DNA repair enzymes are those known as nucleotideexcision repair (NER) enzymes and include those such as Xerodermapigmentosum group C-complementing protein (XPC); RAD23 (S. cerevisiae)homolog (RAD23B); caltractin isoform (CETN2); RFA Protein 1, 2, of 3(RPA1, 2, or 3); 3′ to 5′ DNA helicase (ERCC3); 5′ to 3′ DNA helicase(ERCC2); basic transcription factor (GTF2H1, GTF2H2, GTF2H3, GTF2H4,GTF2H5); CDK activating kinase (CDK7, CCNH); cyclin G1-interactingprotein (MNAT1); DNA excision repair protein ERCC-51; excision repaircross-complementing 1 (ERCC1); DNA ligase 1 (LIG1); ATP-dependenthelicase (ERCC6); and the like.

Also suitable may be DNA repair enzymes in the category that facilitatehomologous recombination and include, but are not limited to DNA repairprotein RAD51 homolog (RAD51, RAD51L1, RAD51B etc.); DNA repair proteinXRCC2; DNA repair protein XRCC3; DNA repair protein RAD52; ATPase(RAD50); 3′ exonuclease (MRE11A); and so on.

DNA repair enzymes that are DNA polymerases are also suitable andinclude DNA polymerase beta subunit (POLB); DNA polymerase gamma (POLG);DNA polymerase subunit delta (POLD1); DNA polymerase II subunit A(POLE); DNA polymerase delta auxiliary protein (PCNA); DNA polymerasezeta (POLZ); MAD2 homolog ((REV7); DNA polymerase eta (POLH): DNApolymerase kappa (POLK): and the like.

Various types of DNA repair enzymes that are often referred to as“editing and processing nucleases” include 3′-nuclease; 3′-exonuclease;5′-exonuclease; endonuclease; and the like.

Other examples of DNA repair enzymes include DNA helicases includingsuch as ATP DNA helicase and so on.

The DNA repair enzymes may be present as components of botanicalextracts, bacterial lysates, biological materials, and the like. Forexample, botanical extracts may contain DNA repair enzymes.

The compositions of the invention may contain one or more DNA repairenzymes.

IV. Other Ingredients

A. Proteasome Activator

The composition may contain one more proteasome activators in amountsranging from about 0.0001 to 65%, preferably from about 0.0005 to 50%,more preferably from about 0.001 to 40%.

Suitable proteasome activators are any compounds, molecules, or activeingredients that stimulate proteasome activity in the cells of keratinsurfaces.

Examples of suitable proteasome activators include, but are not limitedto, algin, alginates, hydrolyzed algin, molasses extract, Trametesextracts, including extracts from Trametes versicolor, olea hydroxol.

The composition of the invention may be in the form of an emulsion,aqueous solution or dispersion, gel, or anhydrous composition. If in theform of an emulsion, it may be a water in oil or oil in water emulsion.If in the form of an emulsion, the composition may contain from about1-99%, preferably from about 5-90%, more preferably from about 10-85%water and from about 1-99%, preferably from about 5-90%, more preferablyfrom about 5-75% of oil. If in the form of an aqueous suspension ordispersion, the composition may generally contain from about 1-99.9%,preferably from about 5-95%, more preferably from about 10-90% water,with the remaining ingredients being the active ingredients or otherformula ingredients.

The composition may additionally contain other ingredients including butnot limited to those set forth herein.

B. CLOCK, PER1 Gene Activator

The composition of the invention may contain a CLOCK or PER1 cellulargene activator. Suggested ranges are from about 0.000001 to about 40%,preferably from about 0.000005 to 35%, more preferably from about0.00001 to 25%. Suitable CLOCK or PER1 activators may be present in theform of botanical extracts, polypeptides, peptides, amino acids, and thelike.

1. Peptide CLOCK or PER1 Gene Activator

A particularly preferred CLOCK and/or PER1 gene activator comprises apeptide of the formula (I):

R₁-(AA)_(n)-X₁-S-T-P—X₂-(AA)_(p)-R₂

Where (AA)_(n)-X₁-S-T—P-X₂-(AA)_(p) is (SEQ ID No. 1) and:

-   -   X₁ represents a threonine, a serine, or is equal to zero,    -   X₂ represents an isoleucine, leucine, proline, valine, alanine,        glycine, or is equal to zero,    -   AA represents any amino acid or derivative thereof, and n and p        are whole numbers between 0 and 4,    -   R₁ represents the primary amine function of the N-terminal amino        acid, either free or substituted by a protective grouping that        may be chosen from either an acetyl group, a benzoyl group, a        tosyl group, or a benzyloxycarbonyl group,        -   R₂ represents the hydroxyl group of the carboxyl function of            the C-terminal amino acid, substituted by a protective            grouping that may be chosen from either a C₁₁ to C₂₀ alkyl            chain or an NH₂, NHY, or NYY group with Y representing a C₁            to C₄ alkyl chain,    -   wherein the sequence of general formula (I) comprises from about        3 to 13 amino acid residues,    -   said sequence of general formula (I) possibly containing        substitutions of amino acids X₁ and X₂ with other chemically        equivalent amino acids;        -   wherein the amino acids are:        -   Alanine (A)        -   Arginine (R)        -   Asparagine (N)        -   Aspartic Acid (D)        -   Cysteine (C)        -   Glutamic Acid (E)        -   Glutamine (Q)        -   Glycine (G)        -   Histidine (H)        -   Isoleucine (I)        -   Leucine (L)        -   Lysine (K)        -   Methionine (M)        -   Phenylalanine (F)        -   Proline (P)        -   Serine (S)        -   Threonine (T)        -   Tryptophan (W)        -   Tyrosine (Y)        -   Valine (V)

More preferred are peptides of the above formula as follows:

S-T-P-NH₂ Ser-Thr-Pro-NH₂ (SEQ ID No. 2) Y-V-S-T-P-Y-N-NH₂Tyr-Val-Ser-Thr-Pro-Tyr-Asn-NH₂ (SEQ ID NO. 3) NH₂-V-S-T-P-E-NH₂NH₂-Val-Ser-Thr-Pro-Glu-NH₂ (SEQ ID No. 4) NH₂-L-H-S-T-P-P-NH₂NH₂-Leu-His-Ser-Thr-Pro-Pro-NH₂ (SEQ ID No. 5) CH₃NH-R-H-S-T-P-E-NH₂CH₃-NH-Arg-His-Ser-Thr-Pro-Glu-NH₂ (SEQ ID No. 6) CH₃NH-H-S-T-P-E-CH₃NHCH₃-NH-His-Ser-Thr-Pro-Glu-CH₃-NH

More preferred is the S-T-P-NH₂ peptide, SEQ ID No. 4, or mixturesthereof.

Most preferred is a peptide manufactured by ISP-Vinscience under thetrademark Chronolux® having the INCI name Tripeptide-32 or Chronogen®having the INCI name Tetrapeptide-26, which has an amino acid sequenceof:

(SEQ ID No. 7) S-P-L-Q-NH₂ Ser-Pro-Leu-Gln-NH₂

2. Botanical Extracts

Also suitable as the CLOCK or PER1 gene activator is cichoric acid orisomers or derivatives thereof. Cichoric acid may be synthetic ornaturally derived. Synthetic cichoric acid may be purchased from anumber of commercial manufacturers including Sigma Aldrich. Cichoricacid may also be extracted from botanical sources that are known tocontain cichoric acid such as Echinacea, Cichorium, Taraxacum, Ocimum,Melissa, or from algae or sea grasses. More specifically, botanicalextracts such as Echinacea purpurea, Cichorium intybus, Taraxacumofficinale, Ocimum basilicum, or Melissa officinalis. The term “cichoricacid” when used herein also includes any isomers thereof that areoperable to increase PER1 gene expression in skin cells.

A specific example includes a botanical extract from Echinacea purpureasold by Symrise under the brand name Symfinity™ 1298 which is an extractof Echinacea purpurea which is standardized during the extractionprocess to contain about 3% by weight of the total extract compositionof cichoric acid. Echinacea extracts from different sources will vary incichoric acid content, and as such will yield variable results ininduction of PER1 gene expression. For example, we have observed thatanother component commonly found in extracts of Echinacea, specificallycaftaric acid, does not increase PER1 gene expression in skin cells.Moreover, each species of Echinacea will differ in content of phenolicand cichoric acids. Ethanolic extract of the roots of Echinacea purpurawill provide more cichoric acid than ethanolic extracts of Echineaceaangustifolia or Echinacea pallida. The content of active ingredients inany extract is also very dependent on the method of extraction. Forexample, it is known that in many cases enzymatic browning during theextraction process will reduce the phenolic acid content of theresulting extract.

C. Sunscreens

It may also be desirable to include one or more sunscreens in thecompositions of the invention. Such sunscreens include chemical UVA orUVB sunscreens or physical sunscreens in the particulate form. Inclusionof sunscreens in the compositions containing the whitening activeingredient will provide additional protection to skin during daylighthours and promote the effectiveness of the whitening active ingredienton the skin. If present, the sunscreens may range from about 0.1 to 50%,preferably from about 0.5 to 40%, more preferably from about 1 to 35%.

1. UVA Chemical Sunscreens

If desired, the composition may comprise one or more UVA sunscreens. Theterm “UVA sunscreen” means a chemical compound that blocks UV radiationin the wavelength range of about 320 to 400 nm. Preferred UVA sunscreensare dibenzoylmethane compounds of the formula:

wherein R₁ is H, OR and NRR wherein each R is independently H, C₁₋₂₀straight or branched chain alkyl; R₂ is H or OH; and R₃ is H, C₁₋₂₀straight or branched chain alkyl.

Preferred is where R₁ is OR where R is a C₁₋₂₀ straight or branchedalkyl, preferably methyl; R₂ is H; and R₃ is a C₁₋₂₀ straight orbranched chain alkyl, more preferably, butyl.

Examples of suitable UVA sunscreen compounds of this general formulainclude 4-methyldibenzoylmethane, 2-methyldibenzoylmethane,4-isopropyldibenzoylmethane, 4-tert-butyldibenzoylmethane,2,4-dimethyldibenzoylmethane, 2,5-dimethyldibenzoylmethane, 4,4′diisopropylbenzoylmethane, 4-tert-butyl-4′-methoxydibenzoylmethane,4,4′-diisopropylbenzoylmethane,2-methyl-5-isopropyl-4′-methoxydibenzoymethane,2-methyl-5-tert-butyl-4′-methoxydibenzoylmethane, and so on.Particularly preferred is 4-tert-butyl-4′-methoxydibenzoylmethane, alsoreferred to as Avobenzone. Avobenzone is commercially available fromGivaudan-Roure under the trademark Parsol® 1789, and Merck & Co. underthe tradename Eusolex® 9020.

Other types of UVA sunscreens include dicamphor sulfonic acidderivatives, such as ecamsule, a sunscreen sold under the trade nameMexoryl®, which is terephthalylidene dicamphor sulfonic acid, having theformula:

The composition may contain from about 0.001-20%, preferably 0.005-5%,more preferably about 0.005-3% by weight of the composition of UVAsunscreen. In the preferred embodiment of the invention the UVAsunscreen is Avobenzone, and it is present at not greater than about 3%by weight of the total composition.

2. UVB Chemical Sunscreens

The term “UVB sunscreen” means a compound that blocks UV radiation inthe wavelength range of from about 290 to 320 nm. A variety of UVBchemical sunscreens exist including alpha-cyano-beta,beta-diphenylacrylic acid esters as set forth in U.S. Pat. No. 3,215,724, which ishereby incorporated by reference in its entirety. One particular exampleof an alpha-cyano-beta,beta-diphenyl acrylic acid ester is Octocrylene,which is 2-ethylhexyl 2-cyano-3,3-diphenylacrylate. In certain cases thecomposition may contain no more than about 10% by weight of the totalcomposition of octocrylene. Suitable amounts range from about 0.001-10%by weight Octocrylene may be purchased from BASF under the tradenameUvinul® N-539.

Other suitable sunscreens include benzylidene camphor derivatives as setforth in U.S. Pat. No. 3,781,417, which is hereby incorporated byreference in its entirety. Such benzylidene camphor derivatives have thegeneral formula:

wherein R is p-tolyl or styryl, preferably styryl. Particularlypreferred is 4-methylbenzylidene camphor, which is a lipid soluble UVBsunscreen compound sold under the tradename Eusolex 6300 by Merck.

Also suitable are cinnamate derivatives having the general formula:

wherein R and R₁ are each independently a C₁₋₂₀ straight or branchedchain alkyl. Preferred is where R is methyl and R₁ is a branched chainC₁₋₁₀, preferably C₈ alkyl. The preferred compound is ethylhexylmethoxycinnamate, also referred to as Octoxinate or octylmethoxycinnamate. The compound may be purchased from GivaudanCorporation under the tradename Parsol® MCX, or BASF under the tradenameUvinul® MC 80.

Also suitable are mono-, di-, and triethanolamine derivatives of suchmethoxy cinnamates including diethanolamine methoxycinnamate. Cinoxate,the aromatic ether derivative of the above compound is also acceptable.If present, the Cinoxate should be found at no more than about 3% byweight of the total composition.

Also suitable as UVB screening agents are various benzophenonederivatives having the general formula:

wherein R through R₉ are each independently H, OH, NaO₃S, SO₃H, SO₃Na,Cl, R″, OR″ where R″ is C₁₋₂₀ straight or branched chain alkyl Examplesof such compounds include Benzophenone 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, and 12. Particularly preferred is where the benzophenone derivativeis Benzophenone 3 (also referred to as Oxybenzone), Benzophenone 4 (alsoreferred to as Sulisobenzone), Benzophenone 5 (Sulisobenzone Sodium),and the like. Most preferred is Benzophenone 3.

Also suitable are certain menthyl salicylate derivatives having thegeneral formula:

wherein R₁, R₂, R₃, and R₄ are each independently H, OH, NH₂, or C₁₋₂₀straight or branched chain alkyl. Particularly preferred is where R₁,R₂, and R₃ are methyl and R₄ is hydroxyl or NH₂, the compound having thename homomethyl salicylate (also known as Homosalate) or menthylanthranilate. Homosalate is available commercially from Merck under thetrademark Eusolex® HMS and menthyl anthranilate is commerciallyavailable from Haarmann & Reimer under the trademark Heliopan®. Ifpresent, the Homosalate should be found at no more than about 15% byweight of the total composition.

Various amino benzoic acid derivatives are suitable UVB absorbersincluding those having the general formula:

wherein R₁, R₂, and R₃ are each independently H, C₁₋₂₀ straight orbranched chain alkyl which may be substituted with one or more hydroxygroups. Particularly preferred is wherein R₁ is H or C₁₋₈ straight orbranched alkyl, and R₂ and R₃ are H, or C₁₋₈ straight or branched chainalkyl. Particularly preferred are PABA, ethyl hexyl dimethyl PABA(Padimate O), ethyldihydroxypropyl PABA, and the like. If presentPadimate O should be found at no more than about 8% by weight of thetotal composition.

Salicylate derivatives are also acceptable UVB absorbers. Such compoundshave the general formula: wherein R is a straight or branched chainalkyl, including derivatives of the above compound formed from mono-,di-, or triethanolamines. Particular preferred are octyl salicylate,TEA-salicylate, DEA-salicylate, and mixtures thereof.

Generally, the amount of the UVB chemical sunscreen present may rangefrom about 0.001-45%, preferably 0.005-40%, more preferably about0.01-35% by weight of the total composition.

If desired, the compositions of the invention may be formulated to havecertain SPF (sun protective factor) values ranging from about 1-50,preferably about 2-45, most preferably about 5-30. Calculation of SPFvalues is well known in the art.

D. Surfactants

It may be desirable for the composition to contain one more surfactants,especially if in the emulsion form. However, such surfactants may beused if the compositions are solutions, suspensions, or anhydrous also,and will assist in dispersing ingredients that have polarity, forexample pigments. Such surfactants may be silicone or organic based. Thesurfactants will also aid in the formation of stable emulsions of eitherthe water-in-oil or oil-in-water form. If present, the surfactant mayrange from about 0.001 to 30%, preferably from about 0.005 to 25%, morepreferably from about 0.1 to 20% by weight of the total composition.

1. Organic Nonionic Surfactants

The composition may comprise one or more nonionic organic surfactants.Suitable nonionic surfactants include alkoxylated alcohols or ethers,formed by the reaction of an alcohol with an alkylene oxide, usuallyethylene or propylene oxide. Suitable alcohols include mono-, di-, orpolyhydric short chain (C1-6) alcohols; aromatic or aliphatic saturatedor unsaturated fatty (C12-40) alcohols, of cholesterol; and so on.

In one embodiment the alcohol is cholesterol, or an aromatic oraliphatic saturated or unsaturated fatty alcohol which may have from 6to 40, preferably from about 10 to 30, more preferably from about 12 to22 carbon atoms. Examples include oleyl alcohol, cetearyl alcohol, cetylalcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, and thelike. Examples of such ingredients include Oleth 2-100; Steareth 2-100;Beheneth 5-30; Ceteareth 2-100; Ceteth 2-100; Choleth 2-100 wherein thenumber range means the number of repeating ethylene oxide units, e.g.Ceteth 2-100 means Ceteth where the number of repeating ethylene oxideunits ranges from 2 to 100. Derivatives of alkoxylated alcohols are alsosuitable, such as phosphoric acid esters thereof.

Some preferred organic nonionic surfactants include Oleth-3, Oleth-5,Oleth-3 phosphate, Choleth-24; Ceteth-24; and so on.

Also suitable are alkoxylated alcohols formed with mono-, di-, orpolyhydric short chain alcohols, for example those having from about 1to 6 carbon atoms. Examples include glucose, glycerin, or alkylatedderivatives thereof. Examples include glycereth 2-100; gluceth 2-100;methyl gluceth 2-100 and so on. More preferred are methyl gluceth-20;glycereth-26 and the like.

Other types of alkoxylated alcohols are suitable surfactants, includingethylene oxide polymers having varying numbers of repeating EO groups,generally referred to as PEG 12 to 200. More preferred are PEG-75, whichis may be purchased from Dow Chemical under the trade name CarbowaxPEG-3350.

Other suitable nonionic surfactants include alkoxylated sorbitan andalkoxylated sorbitan derivatives. For example, alkoxylation, inparticular ethoxylation of sorbitan provides polyalkoxylated sorbitanderivatives. Esterification of polyalkoxylated sorbitan providessorbitan esters such as the polysorbates. For example, thepolyalkyoxylated sorbitan can be esterified with C6-30, preferablyC12-22 fatty acids. Examples of such ingredients include Polysorbates20-85, sorbitan oleate, sorbitan sesquioleate, sorbitan palmitate,sorbitan sesquiisostearate, sorbitan stearate, and so on.

2. Silicone or Silane Surfactants

Also suitable are various types of silicone or silane-based surfactants.Examples include organosiloxanes substituted with ethylene oxide orpropylene oxide groups such as PEG dimethicones which are dimethiconessubstituted with polyethylene glycols including those having the INCInames PEG-1 dimethicone; PEG-4 dimethicone; PEG-8 dimethicone; PEG-12dimethicone; PEG-20 dimethicone; and so on.

Also suitable are silanes substituted with ethoxy groups or propoxygroups or both, such as various types of PEG methyl ether silanes suchas bis-PEG-18 methyl ether dimethyl silane; and so on.

Further examples of silicone based surfactants include those having thegeneric names dimethicone copolyol; cetyl dimethicone copolyol; and soon.

F. Botanical Extracts

It may be desirable to incorporate one more additional botanicalextracts into the composition. If present suggested ranges are fromabout 0.0001 to 20%, preferably from about 0.0005 to 15%, morepreferably from about 0.001 to 10%. Suitable botanical extracts includeextracts from plants (herbs, roots, flowers, fruits, seeds) such asflowers, fruits, vegetables, and so on, including yeast ferment extract,Padina Pavonica extract, Thermus Thermophilis ferment extract, CamelinaSativa seed oil, Boswellia Serrata extract, olive extract, AcaciaDealbata extract, Acer Saccharinum (sugar maple), Acidopholus, Acorus,Aesculus, Agaricus, Agave, Agrimonia, algae, aloe, citrus, Brassica,cinnamon, orange, apple, blueberry, cranberry, peach, pear, lemon, lime,pea, seaweed, caffeine, green tea, chamomile, willowbark, mulberry,poppy, and those set forth on pages 1646 through 1660 of the CTFACosmetic Ingredient Handbook, Eighth Edition, Volume 2. Further specificexamples include, but are not limited to, Glycyrrhiza Glabra, SalixNigra, Macrocycstis Pyrifera, Pyrus Malus, Saxifraga Sarmentosa, VitisVinifera, Morus Nigra, Scutellaria Baicalensis, Anthemis Nobilis, SalviaSclarea, Rosmarinus Officianalis, Citrus Medica Limonum, Panax Ginseng,Siegesbeckia Orientalis, Fructus Mume, Ascophyllum Nodosum, Glycine Sojaextract, Beta Vulgaris, Haberlea Rhodopensis, Polygonum Cuspidatum,Citrus Aurantium Dulcis, Vitis Vinifera, Selaginella Tamariscina,Humulus Lupulus, Citrus Reticulata Peel, Punica Granatum, Asparagopsis,Curcuma Longa, Menyanthes Trifoliata, Helianthus Annuus, HordeumVulgare, Cucumis Sativus, Evernia Prunastri, Evernia Furfuracea, KolaAcuminata, and mixtures thereof. If desired such botanical extracts maybe fermented to increase potency or activity. Fermentation may beaccomplished by standard fermentation techniques using bacteria oryeast.

G. Biological Materials

Also suitable are various types of biological materials such as thosederived from cells, fermented materials, and so on. If present suchmaterials may range from about 0.001 to 30%, preferably from about 0.005to 25%, more preferably from about 0.01 to 20%. Examples includefragments of cellular RNA or DNA, probiotic microorganisms, or fermentsof microorganisms and organic materials from plants such as leaves,seeds, extracts, flowers, etc. Particularly preferred are RNA fragments.

H. Aqueous Phase Structuring Agent

In the case where the compositions are in the form of aqueous solutions,dispersions or emulsions, in addition to water the aqueous phase maycontain one or more aqueous phase structuring agents, that is, an agentthat increases the viscosity or, or thickens, the aqueous phase of thecomposition. This is particularly desirable when the composition is inthe form of a serum or gel. Suitable ranges of aqueous phase structuringagent, if present, are from about 0.01 to 30%, preferably from about 0.1to 20%, more preferably from about 0.5 to 15% by weight of the totalcomposition. Examples of such agents include various acrylate basedthickening agents, natural or synthetic gums, polysaccharides, and thelike, including but not limited to those set forth below.

1. Polysaccharides

Polysaccharides may be suitable aqueous phase thickening agents.Examples of such polysaccharides include naturally derived materialssuch as agar, agarose, alicaligenes polysaccharides, algin, alginicacid, acacia gum, amylopectin, chitin, dextran, cassia gum, cellulosegum, gelatin, gellan gum, hyaluronic acid, hydroxyethyl cellulose,methyl cellulose, ethyl cellulose, pectin, sclerotium gum, xanthan gum,pectin, trehelose, gelatin, and so on.

2. Acrylate Polymers

Also suitable are different types of synthetic polymeric thickeners. Onetype includes acrylic polymeric thickeners comprised of monomers A and Bwherein A is selected from the group consisting of acrylic acid,methacrylic acid, and mixtures thereof; and B is selected from the groupconsisting of a C₁₋₂₂ alkyl acrylate, a C₁₋₂₂ alky methacrylate, andmixtures thereof are suitable. In one embodiment the A monomer comprisesone or more of acrylic acid or methacrylic acid, and the B monomer isselected from the group consisting of a C₁₋₁₀, most preferably C₁₋₄alkyl acrylate, a C₁₋₁₀, most preferably C₁₋₄ alkyl methacrylate, andmixtures thereof. Most preferably the B monomer is one or more of methylor ethyl acrylate or methacrylate. The acrylic copolymer may be suppliedin an aqueous solution having a solids content ranging from about10-60%, preferably 20-50%, more preferably 25-45% by weight of thepolymer, with the remainder water. The composition of the acryliccopolymer may contain from about 0.1-99 parts of the A monomer, andabout 0.1-99 parts of the B monomer. Acrylic polymer solutions includethose sold by Seppic, Inc., under the tradename Capigel.

Also suitable are acrylic polymeric thickeners that are copolymers of A,B, and C monomers wherein A and B are as defined above, and C has thegeneral formula:

wherein Z is —(CH₂)_(m); wherein m is 1-10, n is 2-3, o is 2-200, and Ris a C₁₀₋₃₀ straight or branched chain alkyl. Examples of the secondarythickening agent above, are copolymers where A and B are defined asabove, and C is CO, and wherein n, o, and R are as above defined.Examples of such secondary thickening agents includeacrylates/steareth-20 methacrylate copolymer, which is sold by Rohm &Haas under the tradename Acrysol ICS-1.

Also suitable are acrylate based anionic amphiphilic polymers containingat least one hydrophilic unit and at least one allyl ether unitcontaining a fatty chain. Preferred are those where the hydrophilic unitcontains an ethylenically unsaturated anionic monomer, more specificallya vinyl carboxylic acid such as acrylic acid, methacrylic acid ormixtures thereof, and where the allyl ether unit containing a fattychain corresponds to the monomer of formula:

CH₂═CR′CH₂OB_(n)R

in which R′ denotes H or CH₃, B denotes the ethylenoxy radical, n iszero or an integer ranging from 1 to 100, R denotes a hydrocarbonradical selected from alkyl, arylalkyl, aryl, alkylaryl and cycloalkylradicals which contain from 8 to 30 carbon atoms, preferably from 10 to24, and even more particularly from 12 to 18 carbon atoms. Morepreferred in this case is where R′ denotes H, n is equal to 10 and Rdenotes a stearyl (C18) radical. Anionic amphiphilic polymers of thistype are described and prepared in U.S. Pat. Nos. 4,677,152 and4,702,844, both of which are hereby incorporated by reference in theirentirety. Among these anionic amphiphilic polymers, polymers formed of20 to 60% by weight acrylic acid and/or methacrylic acid, of 5 to 60% byweight lower alkyl methacrylates, of 2 to 50% by weight allyl ethercontaining a fatty chain as mentioned above, and of 0 to 1% by weight ofa crosslinking agent which is a well-known copolymerizable polyethylenicunsaturated monomer, for instance diallyl phthalate,allyl(meth)acrylate, divinylbenzene, (poly)ethylene glycoldimethacrylate and methylenebisacrylamide. One commercial example ofsuch polymers are crosslinked terpolymers of methacrylic acid, of ethylacrylate, of polyethylene glycol (having 10 EO units) ether of stearylalcohol or steareth-10, in particular those sold by the company AlliedColloids under the names SALCARE SC80 and SALCARE SC90, which areaqueous emulsions containing 30% of a crosslinked terpolymer ofmethacrylic acid, of ethyl acrylate and of steareth-10 allyl ether(40/50/10).

Also suitable are acrylate copolymers such as Polyacrylate-3 which is acopolymer of methacrylic acid, methylmethacrylate, methylstyreneisopropylisocyanate, and PEG-40 behenate monomers; Polyacrylate-10 whichis a copolymer of sodium acryloyldimethyltaurate, sodium acrylate,acrylamide and vinyl pyrrolidone monomers; or Polyacrylate-11, which isa copolymer of sodium acryloyldimethylacryloyldimethyl taurate, sodiumacrylate, hydroxyethyl acrylate, lauryl acrylate, butyl acrylate, andacrylamide monomers.

Also suitable are crosslinked acrylate based polymers where one or moreof the acrylic groups may have substituted long chain alkyl (such as6-40, 10-30, and the like) groups, for example acrylates/C₁₀₋₃₀ alkylacrylate crosspolymer which is a copolymer of C10-30 alkyl acrylate andone or more monomers of acrylic acid, methacrylic acid, or one of theirsimple esters crosslinked with the allyl ether of sucrose or the allylether of pentaerythritol. Such polymers are commonly sold under theCarbopol or Pemulen tradenames and have the CTFA name carbomer.

Also suitable are acrylate based polymeric thickeners sold by Clariantunder the Aristoflex trademark such as Aristoflex AVC, which is ammoniumacryloyldimethyltaurate/VP copolymer; Aristoflex AVL which is the samepolymer has found in AVC dispersed in mixture containing caprylic/caprictriglyceride, trilaureth-4, and polyglyceryl-2 sesquiisostearate; orAristoflex HMB which is ammonium acryloyldimethyltaurate/beheneth-25methacrylate crosspolymer, and the like.

3. Alkylene Glycols

Also suitable as the aqueous phase thickening agents are variouspolyethylene glycols (PEG) derivatives where the degree ofpolymerization ranges from 1,000 to 200,000. Such ingredients areindicated by the designation “PEG” followed by the degree ofpolymerization in thousands, such as PEG-45M, which means PEG having45,000 repeating ethylene oxide units. Examples of suitable PEGderivatives include PEG 2M, 5M, 7M, 9M, 14M, 20M, 23M, 25M, 45M, 65M,90M, 115M, 160M, 180M, and the like.

Also suitable are polyglycerins which are repeating glycerin moietieswhere the number of repeating moieties ranges from 15 to 200, preferablyfrom about 20-100. Examples of suitable polyglycerins include thosehaving the CFTA names polyglycerin-20, polyglycerin-40, and the like.

I. Humectants

The composition may contain one or more humectants. If present, they mayrange from about 0.01 to 75%, preferably from about 0.5 to 70%, morepreferably from about 0.5 to 40%. Examples of suitable humectantsinclude glycols, sugars, and the like. Suitable glycols are in monomericor polymeric form and include polyethylene and polypropylene glycolssuch as PEG 4-10, which are polyethylene glycols having from 4 to 10repeating ethylene oxide units; as well as C₁₋₆ alkylene glycols such aspropylene glycol, butylene glycol, pentylene glycol, and the like.Suitable sugars, some of which are also polyhydric alcohols, are alsosuitable humectants. Examples of such sugars include glucose, fructose,honey, hydrogenated honey, inositol, maltose, mannitol, maltitol,sorbitol, sucrose, xylitol, xylose, and so on. Also suitable is urea.Preferably, the humectants used in the composition of the invention areC₁₋₆, preferably C₂₋₄ alkylene glycols, most particularly butyleneglycol.

J. Oils

In the event the compositions of the invention are in emulsion form, thecomposition will comprise an oil phase. Oily ingredients are desirablefor the skin moisturizing and protective properties. Suitable oilsinclude silicones, esters, vegetable oils, synthetic oils, including butnot limited to those set forth herein. The oils may be volatile ornonvolatile, and are preferably in the form of a pourable liquid at roomtemperature. The term “volatile” means that the oil has a measurablevapor pressure, or a vapor pressure of at least about 2 mm. of mercuryat 20° C. The term “nonvolatile” means that the oil has a vapor pressureof less than about 2 mm. of mercury at 20° C. If present, such oils mayrange from about 0.01 to 85%, preferably from about 0.05 to 80%, morepreferably from about 0.1 to 50%.

1. Volatile Oils

Suitable volatile oils generally have a viscosity ranging from about 0.5to 5 centistokes 25° C. and include linear silicones, cyclic silicones,paraffinic hydrocarbons, or mixtures thereof

(a). Volatile Silicones

Cyclic silicones are one type of volatile silicone that may be used inthe composition. Such silicones have the general formula:

where n=3-6, preferably 4, 5, or 6.

Also suitable are linear volatile silicones, for example, those havingthe general formula:

(CH₃)₃Si—O—[Si(CH₃)₂—O]_(n)—Si(CH₃)₃

where n=0, 1, 2, 3, 4, or 5, preferably 0, 1, 2, 3, or 4.

Cyclic and linear volatile silicones are available from variouscommercial sources including Dow Corning Corporation and GeneralElectric. The Dow Corning linear volatile silicones are sold under thetradenames Dow Corning 244, 245, 344, and 200 fluids. These fluidsinclude hexamethyldisiloxane (viscosity 0.65 centistokes (abbreviatedcst)), octamethyltrisiloxane (1.0 cst), decamethyltetrasiloxane (1.5cst), dodecamethyl-pentasiloxane (2 cst) and mixtures thereof, with allviscosity measurements being at 25° C.

Suitable branched volatile silicones include alkyl trimethicones such asmethyl trimethicone having the general formula:

Methyl trimethicone may be purchased from Shin-Etsu Silicones under thetradename TMF-1.5, having a viscosity of 1.5 centistokes at 25° C.

(b). Volatile Paraffinic Hydrocarbons

Also suitable as the volatile oils are various straight or branchedchain paraffinic hydrocarbons having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, or 20 carbon atoms, more preferably 8 to 16 carbonatoms. Suitable hydrocarbons include pentane, hexane, heptane, decane,dodecane, tetradecane, tridecane, and C₈₋₂₀ isoparaffins as disclosed inU.S. Pat. Nos. 3,439,088 and 3,818,105, both of which are herebyincorporated by reference. Preferred volatile paraffinic hydrocarbonshave a molecular weight of 70-225, preferably 160 to 190 and a boilingpoint range of 30 to 320, preferably 60 to 260° C., and a viscosity ofless than about 10 cst. at 25° C. Such paraffinic hydrocarbons areavailable from EXXON under the ISOPARS trademark, and from the PermethylCorporation. Suitable C₁₂ isoparaffins are manufactured by PermethylCorporation under the tradename Permethyl 99A. Various C₁₆ isoparaffinscommercially available, such as isohexadecane (having the tradenamePermethyl R), are also suitable.

2. Non-Volatile Oils

A variety of nonvolatile oils are also suitable for use in thecompositions of the invention. The nonvolatile oils generally have aviscosity of greater than about 5 to 10 centistokes at 25° C., and mayrange in viscosity up to about 1,000,000 centipoise at 25° C. Examplesof nonvolatile oils include, but are not limited to:

(a). Esters

Suitable esters are mono-, di-, and triesters. The composition maycomprise one or more esters selected from the group, or mixturesthereof.

(i). Monoesters

Monoesters are defined as esters formed by the reaction of amonocarboxylic acid having the formula R—COOH, wherein R is a straightor branched chain saturated or unsaturated alkyl having 2 to 45 carbonatoms, or phenyl; and an alcohol having the formula R—OH wherein R is astraight or branched chain saturated or unsaturated alkyl having 2-30carbon atoms, or phenyl. Both the alcohol and the acid may besubstituted with one or more hydroxyl groups. Either one or both of theacid or alcohol may be a “fatty” acid or alcohol, and may have fromabout 6 to 30 carbon atoms, more preferably 12, 14, 16, 18, or 22 carbonatoms in straight or branched chain, saturated or unsaturated form.Examples of monoester oils that may be used in the compositions of theinvention include hexyl laurate, butyl isostearate, hexadecylisostearate, cetyl palmitate, isostearyl neopentanoate, stearylheptanoate, isostearyl isononanoate, steary lactate, stearyl octanoate,stearyl stearate, isononyl isononanoate, and so on.

(ii). Diesters

Suitable diesters are the reaction product of a dicarboxylic acid and analiphatic or aromatic alcohol or an aliphatic or aromatic alcohol havingat least two substituted hydroxyl groups and a monocarboxylic acid. Thedicarboxylic acid may contain from 2 to 30 carbon atoms, and may be inthe straight or branched chain, saturated or unsaturated form. Thedicarboxylic acid may be substituted with one or more hydroxyl groups.The aliphatic or aromatic alcohol may also contain 2 to 30 carbon atoms,and may be in the straight or branched chain, saturated, or unsaturatedform. Preferably, one or more of the acid or alcohol is a fatty acid oralcohol, i.e. contains 12-22 carbon atoms. The dicarboxylic acid mayalso be an alpha hydroxy acid. The ester may be in the dimer or trimerform. Examples of diester oils that may be used in the compositions ofthe invention include diisotearyl malate, neopentyl glycol dioctanoate,dibutyl sebacate, dicetearyl dimer dilinoleate, dicetyl adipate,diisocetyl adipate, diisononyl adipate, diisostearyl dimer dilinoleate,diisostearyl fumarate, diisostearyl malate, dioctyl malate, and so on.

(iii). Triesters

Suitable triesters comprise the reaction product of a tricarboxylic acidand an aliphatic or aromatic alcohol or alternatively the reactionproduct of an aliphatic or aromatic alcohol having three or moresubstituted hydroxyl groups with a monocarboxylic acid. As with themono- and diesters mentioned above, the acid and alcohol contain 2 to 30carbon atoms, and may be saturated or unsaturated, straight or branchedchain, and may be substituted with one or more hydroxyl groups.Preferably, one or more of the acid or alcohol is a fatty acid oralcohol containing 12 to 22 carbon atoms. Examples of triesters includeesters of arachidonic, citric, or behenic acids, such as triarachidin,tributyl citrate, triisostearyl citrate, tri C₁₂₋₁₃ alkyl citrate,tricaprylin, tricaprylyl citrate, tridecyl behenate, trioctyldodecylcitrate, tridecyl behenate; or tridecyl cocoate, tridecyl isononanoate,and so on.

Esters suitable for use in the composition are further described in theC.T.F.A. Cosmetic Ingredient Dictionary and Handbook, Eleventh Edition,2006, under the classification of “Esters”, the text of which is herebyincorporated by reference in its entirety.

(b). Hydrocarbon Oils

It may be desirable to incorporate one or more nonvolatile hydrocarbonoils into the composition. Suitable nonvolatile hydrocarbon oils includeparaffinic hydrocarbons and olefins, preferably those having greaterthan about 20 carbon atoms. Examples of such hydrocarbon oils includeC₂₄₋₂₈ olefins, C₃₀₋₄₅ olefins, C₂₀₋₄₀ isoparaffins, hydrogenatedpolyisobutene, polyisobutene, polydecene, hydrogenated polydecene,mineral oil, pentahydrosqualene, squalene, squalane, and mixturesthereof. In one preferred embodiment such hydrocarbons have a molecularweight ranging from about 300 to 1000 Daltons.

(c). Glyceryl Esters of Fatty Acids

Synthetic or naturally occurring glyceryl esters of fatty acids, ortriglycerides, are also suitable for use in the compositions. Bothvegetable and animal sources may be used.

Examples of such oils include castor oil, lanolin oil, C₁₀₋₁₈triglycerides, caprylic/capric/triglycerides, sweet almond oil, apricotkernel oil, sesame oil, camelina sativa oil, tamanu seed oil, coconutoil, corn oil, cottonseed oil, linseed oil, ink oil, olive oil, palmoil, illipe butter, rapeseed oil, soybean oil, grapeseed oil, sunflowerseed oil, walnut oil, and the like.

Also suitable are synthetic or semi-synthetic glyceryl esters, such asfatty acid mono-, di-, and triglycerides which are natural fats or oilsthat have been modified, for example, mono-, di- or triesters of polyolssuch as glycerin. In an example, a fatty (C₁₂₋₂₂) carboxylic acid isreacted with one or more repeating glyceryl groups. glyceryl stearate,diglyceryl diiosostearate, polyglyceryl-3 isostearate, polyglyceryl-4isostearate, polyglyceryl-6 ricinoleate, glyceryl dioleate, glyceryldiisotearate, glyceryl tetraisostearate, glyceryl trioctanoate,diglyceryl distearate, glyceryl linoleate, glyceryl myristate, glycerylisostearate, PEG castor oils, PEG glyceryl oleates, PEG glycerylstearates, PEG glyceryl tallowates, and so on.

(d). Nonvolatile Silicones

Nonvolatile silicone oils, both water soluble and water insoluble, arealso suitable for use in the composition. Such silicones preferably havea viscosity ranging from about greater than 5 to 800,000 cst, preferably20 to 200,000 cst at 25° C. Suitable water insoluble silicones includeamine functional silicones such as amodimethicone.

For example, such nonvolatile silicones may have the following generalformula:

wherein R and R′ are each independently C₁₋₃₀ straight or branchedchain, saturated or unsaturated alkyl, phenyl or aryl, trialkylsiloxy,and x and y are each independently 1-1,000,000; with the proviso thatthere is at least one of either x or y, and A is alkyl siloxy endcapunit. Preferred is where A is a methyl siloxy endcap unit; in particulartrimethylsiloxy, and R and R′ are each independently a C₁₋₃₀ straight orbranched chain alkyl, phenyl, or trimethylsiloxy, more preferably aC₁₋₂₂ alkyl, phenyl, or trimethylsiloxy, most preferably methyl, phenyl,or trimethylsiloxy, and resulting silicone is dimethicone, phenyldimethicone, diphenyl dimethicone, phenyl trimethicone, ortrimethylsiloxyphenyl dimethicone. Other examples include alkyldimethicones such as cetyl dimethicone, and the like wherein at leastone R is a fatty alkyl (C₁₂, C₁₄, C₁₆, C₁₈, C₂₀, or C₂₂), and the otherR is methyl, and A is a trimethylsiloxy endcap unit, provided such alkyldimethicone is a pourable liquid at room temperature. Phenyltrimethicone can be purchased from Dow Corning Corporation under thetradename 556 Fluid. Trimethylsiloxyphenyl dimethicone can be purchasedfrom Wacker-Chemie under the tradename PDM-1000. Cetyl dimethicone, alsoreferred to as a liquid silicone wax, may be purchased from Dow Corningas Fluid 2502, or from DeGussa Care & Surface Specialties under thetrade names Abil Wax 9801, or 9814.

K. Vitamins and Antioxidants

It may be desirable to incorporate one or more vitamins or antioxidantsin the compositions. If present, suggested ranges are from about 0.001to 20%, preferably from about 0.005 to 15%, more preferably from about0.010 to 10%. Preferably such vitamins, vitamin derivatives and/orantioxidants are operable to scavenge free radicals in the form ofsinglet oxygen. Such vitamins may include tocopherol or its derivativessuch as tocopherol acetate, tocopherol ferulate; ascorbic acid or itsderivatives such as ascorbyl palmitate, magnesium ascorbyl phosphate;Vitamin A or its derivatives such as retinyl palmitate; or vitamins D,K, B, or derivatives thereof.

L. Preferred Compositions

Preferred compositions are in the aqueous solution or emulsion form andcontain at least one autophagy activator and at least one DNA repairenzyme. The composition may optionally contain at least one CLOCK orPER1 gene activator and/or at least one proteasome activator.

More preferred is where within the composition the autophagy activatoris a yeast extract that may be fermented and the DNA repair enzyme isone or more of Arabidopsis thaliana or ferment thereof. If present theproteasome activator is algin, hydrolyzed algin, or alginate. Ifpresent, the nonionic organic surfactant is an alkoxylated alcohol, thechemical sunscreen is a UVB sunscreen, the CLOCK or PER1 keratinocytegene activator is Tripeptide-32 or Tetrapeptide-26, the DNA repairenzyme, if present, is a mixture of Arabidopsis Thaliana extract,Micrococcus lysate, Bifida Ferment lysate, Lactobacillus ferment, andPlankton extract, and the at least one oil is an organic ester orhydrocarbon.

The Method

The invention is also directed to a method for improving normal cellulardetoxification processes by treating the cells with a composition thatstimulates selective catabolysis by activating or enhancing cellularautophagy processes and/or repairing cellular DNA. In the case where aCLOCK or PER1 gene activator may be present, it may improve thesynchronicity of the metabolic pathways of the treated cells, which inturn will improve the efficiency of the autophagy and/or proteasomeactivation processes.

In the method of the invention, the composition may be applied tokeratin surfaces such as skin, scalp, nails or hair one or more timesper day. For example, the composition may be applied to skin in themorning prior to beginning daily activities and/or at night prior toretiring. The composition may be applied as part of a regimen; that is,the skin is cleansed and treated with toner, after which the compositionof the invention is applied. The composition may be part of a kitcontaining a cleanser, toner, and the composition of the invention.

Preferably the composition is applied to the face and/or neck anddécolletage prior to retiring to repair or eliminate damaged cellularmaterial and provide general improvement of the skin. Combining thecomposition of the invention at night prior to retiring maximizes thecellular detoxification processes. In general, treatment of skin withthe composition of the invention promotes cellular viability, longevity,and health.

The invention will be further described in connection with the followingexamples which are set forth for the purposes of illustration only.

Example 1

Normal human dermal fibroblasts were harvested and assayed for cellularviability when untreated, treated with solutions containing 0.0025% ofDNA repair enzyme Arabidopsis thaliana extract, 1% autophagy activatorin the form of yeast extract, and a solution containing a mixture of0.025% Arabidopsis thaliana extract and 1% yeast extract autophagyactivator.

The fibroblasts were placed in concentrations of 150,000 cells per platefor the 48 hour test, and 300,000 cells per plate for the 24 hour teston 96 well plates. The cells were incubated at 37° C., 5% CO₂, and 95%humidity for 24 hours. The test compositions were prepared as follows:

(a) Autophagy Activator.

1% Yeast Extract: 300 ul of yeast extract was added to 14.7 ml of MediaDulbecco's Modified Eagle Media (DMEM) to create a 2% stock solution.Then 2.5 ml of this stock solution was added to 2.5 ml of DMEM to create1% by weight solution.

(b) DNA Repair Enzyme.

Solution of 0.025% Arabidopsis thaliana extract: 15 ul of Roxisomes® wascombined with 14.985 ml of DMEM and filtered to create a 0.1% stocksolution. Then 2.5 ml of this stock solution was diluted with 2.5 ml ofDMEM to create a 0.05% solution. Then 2.5 ml of this 0.05% solution wasdiluted with 2.5 ml of DMEM to make a 0.025% solution of the extract.

(c) Autophagy Activator+DNA Repair Enzyme.

5 ml of the 2% yeast extract solution was mixed with 5 ml of the 0.05%solution of Roxisomes® to form a solution containing 1% yeast extractand 0.025% Roxisomes®.

Normal human dermal fibroblasts were seeded into a 96 well plate at aconcentration of 150,000/well and incubated at 37° C. in 5% CO₂ for 24hours to enable cells to acclimate. The next day each well was treatedwith 200 ul of mixture with each mixture being applied to 6 wells. Theplate was incubated at 37° C. for 72 hours with 5% CO₂.

A 10% solution of Alamar Blue in 100 ul of DMEM was prepared. 100 μl ofa 10% solution of the Alamar Blue reagent was added to each well. Theplate was incubated at 37° C., 5% CO₂ for 2 hours. The fluorescence wasread on a plate reader.

Cells were then irradiated with 10 J/cm² radiation UV Radiation Chamber(Dr. Groebel, UV-Electronik, GmbH). The DPBS was aspirated and thetreatment compositions applied once again for 24 hours. The next morningthe medium was aspirated and 100 μl of 10% Alamar Blue solution wasadded. The plate was incubated at 37° C. for 1.5 to 2 hours. Thefluorescence was measured at 530/590 nanometers using a Spectra MaxGemini reader. The cell viability was calculated and expressed as thepercentage of survival of cells treated with hydrogen peroxide.

1% Yeast Extract + 1% Yeast 0.025% 0.025% Control Extract RoxisomesRoxisomes Average Value 2167.787 2564.93 1798.29 4393.74 Std Dev. 358.51219.54 194.95 308.16 Pvalue 1 0.0008012 0.001132 4.60509E08 % change 018.32 −17.0449 102.68 *expressed as a percentage increase (or decrease,if a negative number) of the untreated control cells.

The above results demonstrate that when cells are treated with thecombination of the autophagy activator and DNA repair enzyme cellularviability is increased 102.68%. Thus, the combination of the autophagyand DNA repair enzyme dramatically improved cellular health andviability both before and after irradiation with UV light.

Example 2

A skin treatment composition in accordance with the invention wasprepared as follows:

Ingredient w/w % Oleth-3 phosphate 0.45 Oleth-3 0.35 Oleth-5 0.24Butylene glycol 0.20 Squalane 0.50 BHT 0.10 Ethylhexyl methoxycinnamate0.10 Choleth-24/ceteth-24 0.10 Triethanolamine 0.11 Retinylpalmitate/zea mays (corn) oil/BHT/BHA 0.10 Butylene glycol 1.1 Chamomile0.03 Bisabolol 0.10 Water QS Methyl paraben 0.46 PEG-75 4.00 Bis-PEG-18methyl ether dimethyl silane 2.00 Glycereth-26 1.00 Methyl gluceth-204.00 Trisodium EDTA 0.10 Pantethine 0.14 Caffeine 0.05 Xanthan gum 0.075Carbomer 0.26 Triethanolamine 0.50 Phenoxyethanol 0.70 Benzyl alcohol0.10 Bifida ferment lysate 9.40 Water/bifida ferment lysate/hydrogenatedlecithin 3.00 Butylene glycol/water/Cola Acuminata extract 3.00 Sodiumribonucleic acid 0.01 Water/butylene glycol/tripeptide-32* 0.20Lactobacillus ferment/lecithin/water 0.05 Water/Arabidopsis Thalianaextract/lecithin**** 0.05 Phenoxyethanol 0.02 Sodium hyaluronate 0.01FD&C Red No. 4 (1% aqueous solution with butylene 0.04 glycol) FD&CYellow No. 5 (1% aqueous solution with butylene 0.09 glycol) D&C GreenNo. 5 (0.1% solution with butylene glycol) 0.001 Yeast extract** 0.001Hydrolyzed Algin (Phyko A1, Codif Recherche & Nature) 0.001 Water QS 100*CLOCK or PER1 gene activator **autophagy activator ***proteasomeactivator ****DNA repair enzyme

The composition was prepared by combining the ingredients and mixingwell to form a liquid. The composition was stored in brown glassbottles.

While the invention has been described in connection with the preferredembodiment, it is not intended to limit the scope of the invention tothe particular form set forth but, on the contrary, it is intended tocover such alternatives, modifications, and equivalents as may beincluded within the spirit and scope of the invention as defined by theappended claims.

We claim:
 1. A topical composition comprising at least one cellularautophagy activator and at least one DNA repair enzyme.
 2. Thecomposition of claim 1 wherein the cells are keratinocytes orfibroblasts.
 3. The composition of claim 1 wherein autophagy activatorcomprises a yeast extract.
 4. The composition of claim 1 wherein theautophagy activator comprises a yeast extract selected from the groupconsisting of those from genuses Lithothamnium, Melilot, Citrus,Candida, Lens, Urtica, Carambola, Momordica, Yarrowia, Plumbago, ormixtures thereof.
 5. The composition of claim 1 wherein the autophagyactivator is selected from the group consisting of Lithothamniumncalcaneum, Melilotus officinalis, Citrus limonum, Candida saitoana, Lensculinaria, Urtica dioica, Averrhoa carambola, Momordica charantia,Yarrowia lipolytica, Plumbago zeylanica.
 6. The composition of claim 1wherein the autophagy activator is a stimulator of one or more of: (a)MAP-LC3; (b) ATG5-12; (c) protein p53; (d) AMPK; or (e) DRAM.
 7. Thecomposition of claim 5 wherein the autophagy activator is an inhibitorof mTOR.
 8. The composition of claim 1 wherein the DNA repair enzyme isselected from the group consisting of base excision repair (BER)enzymes, nucleotide excision repair (NER) enzymes, DNA polymerases, DNAhelicases, Mismatch repair (MMR) enzymes, and mixtures thereof.
 9. Thecomposition of claim 8 wherein the DNA repair enzyme is selected fromthe group consisting of Arabidopsis Thaliana extract either alone or inadmixture with lecithin and water; Lactobacillus ferment, Micrococcuslysate, Plankton extract, Bifida ferment lysate; and mixtures thereof.10. The composition of claim 5 wherein the autophagy activator is yeastextract is from the genus Candida and the DNA repair enzyme comprisesArabidopsis thaliana extract either alone or in admixture with lecithinand water.
 11. The composition of claim 1 additionally comprising atleast one proteasome activator.
 12. The composition of claim 11 whereinthe proteasome activator is algin, alginate, or hydrolyzed algin. 13.The composition of claim 1 additionally comprising a CLOCK or PER1 geneactivator comprising: (a) a peptide having from about 3 to 13 amino acidresidues and of the formula (I) (SEQ ID No. 1):R₁-(AA)_(n)-X₁-S-T-P-X₂-(AA)_(p)-R₂, wherein X₁ represents threonine,serine, or is equal to zero, X₂ represents an isoleucine, leucine,proline, valine, alanine, glycine, or is equal to zero, AA representsany amino acid or derivative thereof, and n and p are whole numbersbetween 0 and 4, R₁ represents the primary amine function of theN-terminal amino acid, either free or substituted by a protectivegrouping that may be chosen from either an acetyl group, a benzoylgroup, a tosyl group, or a benzyloxycarbonyl group, R₂ represents thehydroxyl group of the carboxyl function of the C-terminal amino acid,which may be substituted by a protective grouping that may be chosenfrom either a C₁ to C₂₀ alkyl chain or an NH₂, NHY, or NYY group with Yrepresenting a C₁ to C₄ alkyl chain, and wherein said formula (I)sequence may contain substitutions of amino acids X₁ and X₂ with otherchemically equivalent amino acids; (b) a peptide having the amino acidsequence Ser-Pro-Leu-Gln-NH₂; (c) cichoric acid; and (d) mixturesthereof.
 14. The composition of claim 13 wherein the CLOCK or PER1 geneactivators are selected from the group consisting of: S-T-P-NH₂Ser-Thr-Pro-NH₂ (SEQ ID No. 2) Y-V-S-T-P-Y-N-NH₂Tyr-Val-Ser-Thr-Pro-Tyr-Asn-NH₂ (SEQ ID No. 3) NH₂-V-S-T-P-E-NH₂NH₂-Val-Ser-Thr-Pro-Glu-NH₂ (SEQ ID No. 4) NH₂-L-H-S-T-P-P-NH₂NH₂-Leu-His-Ser-Thr-Pro-Pro-NH₂ (SEQ ID No. 5) CH₃NH-R-H-S-T-P-E-NH₂CH₃-NH-Arg-His-Ser-Thr-Pro-Glu-NH₂ (SEQ ID No. 6) CH₃NH-H-S-T-P-E-CH₃NHCH₃-NH-His-Ser-Thr-Pro-Glu-CH₃-NH₂

and mixtures thereof.
 15. The composition of claim 13 wherein the CLOCKor PER1 gene activators are selected from: S-T-P-NH₂ Ser-Thr-Pro-NH₂(SEQ ID No. 4) NH₂-L-H-S-T-P-P-NH₂ NH₂-Leu-His-Ser-Thr-Pro-Pro-NH₂

and mixtures thereof.
 16. The composition of claim 1 additionallycomprising a CLOCK or PER1 gene activator selected from the groupconsisting of Tripeptide-32, Tetrapeptide-26 or mixtures thereof.
 17. Amethod for increasing cellular viability and selective catabolysis incells of keratin surfaces in need of such treatment comprising applyingto the surface a composition comprising at least one autophagy activatorand at least one DNA repair enzyme.
 18. The method of claim 17 whereinthe cellular viability and selective autophagy activation results inimproved cellular detoxification.
 19. The method of claim 17 wherein thecomposition is applied at night prior to evening rest.
 20. The method ofclaim 17 wherein the keratin surfaces are skin, hair, or nails.